Unconfigured Ad

Collapse
X
 
  • Time
  • Show
Clear All
new posts
  • nurmp89
    Junior Member
    • Jan 2013
    • 1

    #1

    Sanger sequencing chromatograms will look like what for heterozygous mutation?

    Hello,

    I have a question: For heterozygous deletion, Sanger sequencing chromatograms will look like what?

    This is because, I found heterozygous deletion in 1 sample when i did ion torrent sequencing. Brief information about that mutation is below with IGV figure.

    Chr Start End Ref Alt Func.refGene Gene.refGene
    chr1 155208272 155208276 AGGGC AGGC intronic GBA

    Click image for larger version

Name:	PD272 Ion Torrent - IGV.png
Views:	1
Size:	97.2 KB
ID:	308968Click image for larger version

Name:	PD272 Ion Torrent - IGV 2.PNG
Views:	1
Size:	4.1 KB
ID:	308970

    I validate this mutation using Sanger sequencing and I got this result. Does this result correct? Does it is represent heterozygous deletion?

    Click image for larger version

Name:	PD272 Sanger - Reverse.PNG
Views:	1
Size:	42.0 KB
ID:	308969
  • Bukowski
    Senior Member
    • Jan 2010
    • 388

    #2
    Normally a het indel in a Sanger trace will mean you end up with superimposed traces. See:

    Direct Sanger sequencing of a diploid template containing a heterozygous insertion or deletion results in a difficult-to-interpret mixed trace formed by two allelic traces superimposed onto each other. Existing computational methods for ...


    You don't actually show the read support in IGV in your screenshots, but from the pop up it looks pretty minimal. Looks like the reference sequence to me.
    Last edited by Bukowski; 02-04-2016, 02:51 AM.

    Comment

    • sklages
      Senior Member
      • May 2008
      • 628

      #3
      Originally posted by nurmp89 View Post
      Hello,
      I validate this mutation using Sanger sequencing and I got this result. Does this result correct? Does it is represent heterozygous deletion?
      [ATTACH]4185[/ATTACH]
      Usually for heterozygous indels the sequence after the start of the mutation is more or less "unreadable" or hard to read due to both alleles being read by the polymerase.

      See example here http://www.codoncode.com/aligner/quicktour/indels.htm

      best,
      Sven

      Comment

      Latest Articles

      Collapse

      • SEQadmin2
        16S/ITS Profiling vs. Shotgun Metagenomics: What Changes in Your Workflow?
        by SEQadmin2




        NGS has revolutionized clinical microbiology by offering new ways to identify and surveil pathogens without prior knowledge of the target. One popular approach is 16S rRNA and ITS amplicon sequencing, which targets specific microbial markers. But studies are increasingly incorporating shotgun metagenomics, a technique that sequences all DNA from a sample without targeting specific microbial markers. This approach allows more detailed taxonomic classification, detection...
        Yesterday, 06:46 AM
      • SEQadmin2
        Multiomic Liquid Biopsy: What DNA/RNA Panels Reveal That DNA Alone Can’t
        by SEQadmin2



        Tumors contain complex mixtures of cell populations that evolve in response to treatment and host-specific factors. Tissue biopsies provide only static snapshots of a tumor’s state. But what if we could follow that evolution over time using a minimally invasive technique?

        Liquid biopsies can detect tumor-derived material circulating in the bloodstream, providing a way to study treatment response, detect emerging resistance, and discover new biomarkers. Circulating
        ...
        10-14-2026, 09:55 AM

      ad_right_rmr

      Collapse

      News

      Collapse

      Topics Statistics Last Post
      Started by SEQadmin2, 10-28-2026, 09:57 AM
      0 responses
      25 views
      0 reactions
      Last Post SEQadmin2  
      Started by SEQadmin2, 10-28-2026, 09:51 AM
      0 responses
      15 views
      0 reactions
      Last Post SEQadmin2  
      Started by SEQadmin2, 10-22-2026, 11:13 AM
      0 responses
      71 views
      0 reactions
      Last Post SEQadmin2  
      Started by SEQadmin2, 10-22-2026, 10:48 AM
      0 responses
      48 views
      0 reactions
      Last Post SEQadmin2  
      Working...